Free Testosterone and Free T3/T4

Measurement of FT4 and FT3 is typically performed by immunoassay, however these can be prone to suffer from interferences and a lack of selectivity. Currently, the accepted gold standard for the measurement of FT4 and FT3 is using equilibrium dialysis (ED) and combined with the use of LC-MS/MS can overcome the issues with immunoassay in the measurement of FT4 and FT3. However, the equilibrium dialysis procedure can be quite time consuming and often being performed overnight.

Here, we describe a simple clinical research method to perform ED using a commercially available device followed by sample preparation by solid phase extraction (SPE) and UPLC-MS/MS analysis using the Waters™ ACQUITY™ UPLC and analytically sensitive Xevo™ TQ Absolute Mass Spectrometer for clinical research. Data analysis and review has been simplified with the use of the waters_connect™ with QUAN Review Software. This has allowed for the complete workflow including equilibrium dialysis to be performed within a typical working day.

Simplified Equilibrium Dialysis for the UPLC-MS/MS Analysis of Free Testosterone (FT) for Clinical Research

Historically, the measurement of free testosterone in human serum has presented a challenge for laboratory scientists, with such a large portion bound and the complexity and time involved in “gold standard” equilibrium dialysis. With a simple, fast, workflow combined with the analytical sensitivity afforded by the Waters Xevo™ TQ Absolute Mass Spectrometer, a newly developed, precise, accurate and selective clinical research method will be presented.

Presenter: Robert Wardle (Senior Clinical Scientist, Waters Corporation)

Robert Wardle is currently a Senior Scientist employed at Waters Corporation in Wilmslow, UK, performing research and development of applications and products for Clinical Research. His primary area of research is vitamins and in particular vitamin D metabolites, steroid hormones and in-depth knowledge of offline and online automation.

Robert joined Waters in 2010 as a Clinical Validation Scientist, playing a key role in verification activities for the MassTrak XE Immunosuppressants kit available in the EU. He moved into his current role as a Senior Scientist in 2013, where he developed clinical research methods for a range of applications including vitamin D metabolites, estrogens, a large panel of steroid hormones and more recently this application on free T4 and free T3.

Presenter: Stephen Balloch (Senior Clinical Scientist, Waters Corporation)

Stephen Balloch graduated from the University of York (UK) with a Master’s degree in Chemistry, which included a strong focus on analytical chemistry. Subsequently he worked as a laboratory scientist in a GLP compliant research facility, progressing to Study Director. And, since 2010, he has been part of the clinical team at Waters (Wilmslow, UK). His principal focus has been on the development and validation of bioanalytical methods according to CLSI guidelines, with a focus on therapeutic drug monitoring. Additionally, he has supported newborn screening, vitamin and steroid analysis, including this clinical research method for free testosterone